Vaccinia virus is used to vaccinate persons to prevent disease with smallpox. Compared with other live virus vaccines, the smallpox vaccine is associated with frequent side effects including fever, a sore or swollen arm, headache, and fatigue. Other more serious side effects have also been reported including development of multiple lesions outside the vaccination site (generalized vaccinia), inflammation of the brain (postvaccinia encephalitis), severe infection of the skin in persons with a history of eczema (eczema vaccinatum), and disseminated vaccinia virus infection in immunocompromised persons (progressive vaccinia). Previously, we tested blood from smallpox vaccine recipients who received the Dryvax formulation of vaccine and found that 4 of 202 blood samples (from 3 of 27 vaccine recipients) were positive for the smallpox vaccine virus in the blood by PCR, but none were positive by culture. The lack of finding infectious smallpox vaccine virus in the blood indicates that the current guidelines for deferral of blood donation from smallpox vaccine recipients are appropriate. We tested throat swabs and found that 0 of 213 throat swabs were positive by culture and 0 of 190 throat swabs were positive by PCR. This study suggests that few, if any, healthy smallpox vaccine recipients shed the virus from their throat. Since the study was begun, the Dryvax formulation of smallpox vaccine has been replaced by the ACAMBIS 2000 formulation. We are now testing persons who received the ACAMBIS 2000 formulation of vaccinia and will be determining if blood samples and throat swabs might contain virus.